Abstract
Growth arrest specific (gas) 1 gene product is expressed in non-transformed fibroblasts in response to stimuli driving cells into Go phase. Gas1 has been demonstrated to inhibit cell proliferation when over-expressed in proliferating fibroblasts. This activity depends on a function of the p53 protein independent of its transactivating ability. To better define the pathway leading from Gas1, which is located on the plasma membrane, to p53, we have undertaken a detailed characterization of its topology. We demonstrate that the protein undergoes cotranslational modifications in the endoplasmic reticulum, consisting of signal peptide cleavage, N-linked glycosylation and glycosyl-phosphatidylinositol anchor addition. Immunoelectron microscopy shows that, in its mature form, Gas1 is randomly distributed over the outer leaflet of the plasma membrane and that upon antibody-induced clustering it relocalizes to caveolae.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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3T3 Cells
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Animals
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COS Cells
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Cell Cycle Proteins
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Cell Division
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Cell Membrane / metabolism*
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Consensus Sequence / physiology
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Endoplasmic Reticulum / metabolism
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GPI-Linked Proteins
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Glutaral
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Glycosylphosphatidylinositols / metabolism*
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Humans
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Membrane Glycoproteins / chemistry
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Membrane Glycoproteins / genetics
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Membrane Glycoproteins / metabolism*
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Membrane Glycoproteins / ultrastructure
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Membrane Proteins
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Mice
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Microscopy, Immunoelectron
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Palmitic Acid / metabolism
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Phosphatidylinositol Diacylglycerol-Lyase
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Precipitin Tests
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Protein Binding
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Protein Sorting Signals / physiology
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Saccharomyces cerevisiae Proteins*
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Tissue Fixation
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Transfection
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Type C Phospholipases / metabolism
Substances
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Cell Cycle Proteins
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GAS1 protein, S cerevisiae
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GAS1 protein, human
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GPI-Linked Proteins
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Gas1 protein, mouse
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Glycosylphosphatidylinositols
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Membrane Glycoproteins
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Membrane Proteins
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Protein Sorting Signals
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Saccharomyces cerevisiae Proteins
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Palmitic Acid
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Type C Phospholipases
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Phosphatidylinositol Diacylglycerol-Lyase
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Glutaral